CN112844470A - Preparation method and application of magnetic silicon-based functional nano enzyme material - Google Patents
Preparation method and application of magnetic silicon-based functional nano enzyme material Download PDFInfo
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- 239000000463 material Substances 0.000 title claims abstract description 60
- 229910052710 silicon Inorganic materials 0.000 title claims abstract description 51
- 239000010703 silicon Substances 0.000 title claims abstract description 51
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 39
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000002122 magnetic nanoparticle Substances 0.000 claims abstract description 18
- 239000002245 particle Substances 0.000 claims abstract description 15
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000002253 acid Substances 0.000 claims abstract description 10
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 10
- 235000012239 silicon dioxide Nutrition 0.000 claims abstract description 7
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- 238000003756 stirring Methods 0.000 claims description 36
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- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 10
- 239000011324 bead Substances 0.000 claims description 10
- 239000001509 sodium citrate Substances 0.000 claims description 10
- 239000011550 stock solution Substances 0.000 claims description 10
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 10
- 229940038773 trisodium citrate Drugs 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 claims description 9
- 239000005457 ice water Substances 0.000 claims description 9
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 9
- 239000012279 sodium borohydride Substances 0.000 claims description 9
- 239000007787 solid Substances 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 8
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 7
- 238000005576 amination reaction Methods 0.000 claims description 7
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 7
- 229910052737 gold Inorganic materials 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 238000007885 magnetic separation Methods 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 235000017281 sodium acetate Nutrition 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 238000007865 diluting Methods 0.000 claims description 4
- 235000015243 ice cream Nutrition 0.000 claims description 4
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 claims description 3
- 239000003054 catalyst Substances 0.000 claims description 3
- QKUSRAKPUWQSJS-UHFFFAOYSA-N diazanium 3-ethyl-2H-1,3-benzothiazole-6-sulfonate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)C1=CC=C2N(CC)CSC2=C1.[O-]S(=O)(=O)C1=CC=C2N(CC)CSC2=C1 QKUSRAKPUWQSJS-UHFFFAOYSA-N 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000012546 transfer Methods 0.000 claims description 3
- 230000032683 aging Effects 0.000 claims description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 14
- 230000003197 catalytic effect Effects 0.000 abstract description 10
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 abstract description 7
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 5
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- 230000007547 defect Effects 0.000 abstract description 3
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- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 229910052697 platinum Inorganic materials 0.000 abstract 1
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
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- 239000011259 mixed solution Substances 0.000 description 4
- 239000002105 nanoparticle Substances 0.000 description 4
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- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 238000001132 ultrasonic dispersion Methods 0.000 description 3
- 238000003917 TEM image Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 229910052681 coesite Inorganic materials 0.000 description 2
- 229910052906 cristobalite Inorganic materials 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 229910052682 stishovite Inorganic materials 0.000 description 2
- 229910052905 tridymite Inorganic materials 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
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- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 229910052976 metal sulfide Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
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Images
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- B01J35/33—
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/003—Catalysts comprising hydrides, coordination complexes or organic compounds containing enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/02—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides
- B01J31/0272—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides containing elements other than those covered by B01J31/0201 - B01J31/0255
- B01J31/0274—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides containing elements other than those covered by B01J31/0201 - B01J31/0255 containing silicon
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/02—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides
- B01J31/0272—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides containing elements other than those covered by B01J31/0201 - B01J31/0255
- B01J31/0275—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides containing elements other than those covered by B01J31/0201 - B01J31/0255 also containing elements or functional groups covered by B01J31/0201 - B01J31/0269
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/26—Catalysts comprising hydrides, coordination complexes or organic compounds containing in addition, inorganic metal compounds not provided for in groups B01J31/02 - B01J31/24
- B01J31/28—Catalysts comprising hydrides, coordination complexes or organic compounds containing in addition, inorganic metal compounds not provided for in groups B01J31/02 - B01J31/24 of the platinum group metals, iron group metals or copper
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- B01J35/39—
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- B01J35/393—
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- B01J35/40—
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
Abstract
The invention belongs to the field of catalytic materials, and particularly relates to a preparation method and application of a magnetic silicon-based functional nano enzyme material. The surface of the magnetic nano-particles is modified with silicon dioxide and aminated, so that nano-platinum particles are loaded, and the bimetallic nano-enzyme material with high catalytic activity is formed. The functional nano enzyme material not only has the characteristics of natural enzyme, but also can make up the defects of the natural enzyme, such as: the color developing agent has the characteristics of good stability, low cost, acid resistance, reusability, easy recovery and the like, can be used for controllably modifying and modifying, can be used for developing ABTS, OPD and TMB without adding hydrogen peroxide, and greatly saves the reagent cost. Fe3O4Introduction of magnetic nanoparticlesThe nano enzyme has the characteristic of easy separation, and can be separated and recovered without the help of a high-speed centrifuge, thereby greatly widening the application field of the nano enzyme.
Description
Technical Field
The invention belongs to the field of catalytic materials, and particularly relates to a preparation method and application of a magnetic silicon-based functional nano enzyme material.
Background
The natural enzyme is used as an efficient green catalyst and has important application in the fields of food, environment, biology, chemical industry and the like. However, in practical application, natural enzymes are limited by a plurality of environmental factors such as pH, temperature, metal ions, time and the like, so that the natural enzymes are volatile and have poor stability, and have the defects of high purification cost, difficult recovery, high price and the like. Therefore, the development of nano enzyme materials is expected to solve the existing problems of natural enzymes.
The nano-enzyme material has been widely studied because of its unique physicochemical properties and its similarity to the size, shape, catalytic activity, etc. of natural enzymes. The nano enzyme material has the advantages of low cost, good stability, reusability, high catalytic efficiency and the like, has controllable composition and structure, can be modified and modified, and has great interest for researchers in recent years.
In recent years, research work on nanoenzyme materials has been increasing, for example: metal nano-particles, metal oxide nano-particles, metal sulfides, carbon-based nano-materials and the like, wherein the bimetallic nano-enzyme is widely used in the field of catalysis, and magnetic Fe is added on the basis3O4The enzyme is easy to separate and recycle, the operation steps are simplified, and the purpose of portability is achieved.
Disclosure of Invention
Aiming at the defects of the existing nano enzyme material, the invention aims to provide a preparation method and application of a magnetic silicon-based functional nano enzyme material, the material can enable ABTS, OPD and TMB to develop color, hydrogen peroxide is not added, and the material has the advantages of low cost, good stability, high catalytic activity, acid resistance, easiness in recovery and the like; and the method is easy to separate and synthesize, and is beneficial to practical popularization and application.
The invention is realized by the following technical scheme:
1. a preparation method of a magnetic silicon-based functional nano enzyme material comprises the following reagents:
A. ferric chloride (FeCl)3)；
B. Trisodium citrate (Na)3C6H5O7·2H2O)；
C. Chloroauric acid (HAuCl)4，24.32mM)；
D. Sodium borohydride (NaBH)4)；
E. Ethylene glycol;
F. sodium acetate;
G. ammonia (NH)3·H2O，25％)
H. Tetraethyl orthosilicate (TEOS, 99%);
I. (3-aminopropyl) triethoxysilane (APTES, 97%);
J. chloroplatinic acid (H)2PtCl6,19.3mM)；
k. 3,3 ', 5, 5' -tetramethylbenzidine (TMB, 1.25 mM);
l, OPD (o-phenylenediamine);
m, ABTS (2, 2-diaza-bis (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt).
Wherein
The method comprises the following steps:
1. synthesis of gold nanoparticles (AuNPS): taking 0.1-3 mL of 15-30 mM chloroauric acid and 4-10 g of trisodium citrate, diluting to a constant volume of 100mL, then placing on a magnetic stirrer, and adding 0.5-5 mLNaBH4And stirring for 10-12 h at normal temperature.
Preferably, NaBH4The ice cream is prepared by dissolving 5-20 mg of the ice cream in 1-5 mL of ice water.
2、Fe3O4Magnetic nanoparticle synthesis: 0.5g to 10g of FeCl3And 0.1-2 g of trisodium citrate are dispersed in 20-40 mL of ethylene glycol, then 0.1-3 g of sodium acetate is added under the stirring condition, the stirring is continued for 20-60 min, the mixture is transferred into a reaction kettle, the reaction is carried out for 8-12 h under the condition of 180-300 ℃, then the reaction is cooled to room temperature, and the mixture is washed with water and ethanol for multiple times, preferably for 5 times.
3、Fe3O4Magnetic nano-meterCoating silicon dioxide on the surface of the particle: and (3) ultrasonically dispersing 100-800 mu L of synthetic magnetic bead stock solution into 5mL of ultrapure water, then sucking 5-35 mL of absolute ethyl alcohol by using a liquid transfer gun, adding the absolute ethyl alcohol into a three-neck flask, after ultrasonic dispersion is carried out for 2-5 min, erecting the three-neck flask and a stirring device at the rotating speed of 300-500 rpm (sufficient stirring and medium speed), then adding 100-500 mu L of concentrated ammonia water into the three-neck flask, and stirring for 5-10 min. At this time, 10 to 100. mu.L of TEOS solution was added dropwise, and the mixture was stirred at room temperature for reaction overnight. After the reaction is finished, cleaning the reaction product with ethanol and ultrapure water for three times, and finally adding water to a constant volume of 1-10 mL. Wherein the concentrated ammonia is added slowly.
Preferably, the TEOS solution is prepared by mixing TEOS and absolute ethanol in a ratio of 1: 1.
4. Amination: taking 100-800 mu L of stock solution coated with silicon, using ethanol to fix the volume to 5mL, then adding APTES solution, fully mixing, adding 20-200 mu L of concentrated ammonia water, stirring at room temperature to react for 6-20 h, after the reaction is finished, washing with ethanol for 3 times, and fixing the volume to 2 mL.
Preferably, the APTES solution is prepared by dissolving 0.5-25 uL of APTES in 50-650 uL of ethanol, and adding the APTES solution completely.
5. And taking 10-800 mu L of aminated magnetic nanoparticles, removing ethanol, adding Au NPs, performing ultrasonic treatment for 15min, performing magnetic separation to obtain a supernatant with transparent color and no AuNPs color, judging that the Au NPs are successfully adsorbed on the surface of the magnetic beads, and repeating the process until the supernatant is wine red in color. Re-dispersing the obtained magnetic particles loaded with the nanogold in 2-10 mL of ultrapure water, and slowly adding 50-500 mu L of H with the concentration of 10-25 mM under the stirring condition2PtCl6After reacting for 5-20 min, slowly adding NaBH4And aging the solution for 3-12 h, finally carrying out centrifugal cleaning, retaining the solid, re-dispersing and storing.
Preferably, NaBH4The solution is prepared by dissolving 0.001-0.1 g of the solution in 1-5 mL of ice water, and adding the solution completely.
6. And (3) putting 0.5-1.5 mL of the prepared functional magnetic nano enzyme material into a glass bottle, adding 50-200 mu L of TMB, standing for 1-5 min, and enabling the mixed solution to show blue.
Any one of the magnetic silicon-based functional nano enzyme materials is used for the color development of 3,3 ', 5, 5' -tetramethyl benzidine, o-phenylenediamine or 2, 2-diazo-bis (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt.
The invention has the beneficial effects that: the magnetic silicon-based functional nano enzyme material provided by the invention has the following characteristics: 1. the high catalytic performance of natural enzyme, the unique optical property of Au and the excellent catalytic property of Pt are considered; 2. TMB, OPD or ABTS can be developed without adding hydrogen peroxide, so that the reagent cost is greatly reduced, and the color can be developed more quickly by adding hydrogen peroxide; 3. acid resistance and stability under acidic conditions; 4. the magnetic material is magnetic and easy to recycle; 5. repeated use, stable property and the like, and is beneficial to practical popularization and application.
Drawings
FIG. 1: a synthetic process diagram of the magnetic silicon-based functional nano enzyme material;
FIG. 2: a particle size diagram of the magnetic particles, the magnetic particle-loaded nanogold and the magnetic silicon-based functional nanoenzyme material of example 1;
FIG. 3: zeta graphs of the magnetic particle, the magnetic particle silicon-coated ammoniated and magnetic silicon-based functional nanoenzyme materials of the embodiment 1;
FIG. 4: a magnet separation effect diagram of the magnetic particles, the magnetic particle-loaded nanogold and the magnetic silicon-based functional nanoenzyme material of example 1;
FIG. 5: TEM images of the magnetic particles, magnetic particle-silicon-coated and magnetic silicon-based functional nanoenzyme materials of example 1;
FIG. 6: the magnetic silicon-based functional nano enzyme material in the embodiment 1 catalyzes TMB to develop color;
FIG. 7: a catalytic effect diagram of the magnetic silicon-based functional nanoenzyme material of example 1 under different pH values;
FIG. 8: the color development effect graphs of the magnetic silicon-based functional nano-enzyme material of the embodiment 1 on ABTS, OPD and TMB respectively.
Detailed Description
Example 1
A preparation method of a magnetic silicon-based functional nano enzyme material comprises the following steps:
1. nano gold particles (AuN)PS) synthesis: 2mL of 16mM chloroauric acid and 7.3g of trisodium citrate are taken, the volume is adjusted to 100mL, then the mixture is placed on a magnetic stirrer, and 3mL of NaBH is added4(taking 11mgNaBH4Dissolved in 3mL of ice water), and stirred at room temperature overnight.
2、Fe3O4Magnetic nanoparticle synthesis: 0.5g of FeCl3And 0.2g trisodium citrate in 25mL of ethylene glycol, then adding 1.2g sodium acetate under stirring, continuously stirring for 30min, transferring into a reaction kettle, reacting at 180 ℃ for 12h, then cooling to room temperature, washing with water and ethanol for 5 times. 3. Fe3O4Coating silicon dioxide on the surface of the magnetic nano-particles: and (3) ultrasonically dispersing 500 mu L of the synthesized stock solution of the magnetic beads in 5mL of ultrapure water, sucking 25mL of absolute ethyl alcohol by using a liquid transfer gun, adding the absolute ethyl alcohol into a three-neck flask, after ultrasonic dispersion is carried out for 5min, erecting the three-neck flask and a stirring device at the rotating speed of 300-500 rpm (sufficient stirring and medium speed), adding 200 mu L of concentrated ammonia water (slowly adding), and stirring for 5-10 min. At this time, 50 μ L of TEOS (TEOS: absolute ethanol ═ 1:1) was further added dropwise, and the reaction was stirred at room temperature overnight. After the reaction is finished, the solution is washed with ethanol and ultrapure water for three times, and finally the volume is adjusted to 5mL by water.
4. Amination: taking 500 mu L of stock solution coated with silicon, using ethanol to fix the volume to 5mL, then adding 16 mu L of LAPTES (dissolved in 500 mu L of ethanol), fully mixing, adding 125 mu L of strong ammonia water, stirring at room temperature for reaction for 12h, finishing the reaction, washing with ethanol for 3 times, and fixing the volume to 2 mL.
5. And (3) taking 500 mu L of aminated magnetic nanoparticles, removing ethanol, adding Au NPs, performing ultrasonic treatment for 15min, performing magnetic separation to obtain a supernatant with transparent color and no Au NPs color, judging that the Au NPs are successfully adsorbed on the surfaces of the magnetic beads, and repeating the process until the supernatant is wine red in color. Re-dispersing the obtained magnetic particles loaded with the nanogold in 2-10 mL of ultrapure water, and slowly adding 400 mu L H under the stirring condition2PtCl6(19.32mM), reacting for 5-20 min, and slowly adding NaBH4(0.017 g of the solid is dissolved in 2mL of ice water), then the mixture is aged for 12 hours, and finally the solid is retained after centrifugal cleaning, and the solid is stored after redispersion.
6. Putting 0.7mL of the prepared functional magnetic nano material into a glass bottle, adding 200 μ L of TMB, and standing for 1.5min to obtain the picture 6. Left in fig. 6: magnetic bead function nanoenzyme, right: magnetic bead functional nanoenzyme + TMB; as can be seen from FIG. 6, the mixed solution appeared blue, which indicated that the prepared nanoenzyme material had the function of catalyzing the color development of TMB, and no hydrogen peroxide was added.
FIG. 2 is a particle size diagram of the magnetic particles, magnetic-particle-loaded nano-gold and magnetic silicon-based functional nano-enzyme material of the present embodiment; the particle size of the synthesized magnetic nanoparticles is 197.3nm through the measurement of a dynamic light scattering instrument, the particle size is uniform, and the dispersibility is good; after silica is wrapped and aminated, a layer of nano gold particles is loaded on the surfaces of the magnetic particles through electrostatic adsorption, and the particle size is increased from 197.3nm to 308.4 nm; and then the particle size is increased to 741nm, which shows that Pt is successfully covered on MB @ Au, namely the functional nano enzyme material is successfully synthesized.
FIG. 3 is a zeta diagram of the magnetic particle, the silicon-coated magnetic particle, the ammoniated and magnetic silicon-based functional nanoenzyme material after the silicon-coated magnetic particle is coated with silicon in the embodiment; wrapped with SiO2The surface potential of the magnetic particle is-37.2 mV, the potential is increased to +18.1mV after amination, and the potential is changed to-14.5 mV after PtAu is loaded, which can indicate that PtAu is successfully modified on the surface of the magnetic particle.
FIG. 4 is a diagram showing the effect of magnetic separation of magnetic particles, magnetic-particle-loaded nanogold and magnetic silicon-based functional nanoenzyme material according to the embodiment; the left side is before magnet separation, and the right side is after magnet separation, and as can be seen from the figure, the three samples after magnet separation are colorless, which proves that the prepared magnetic silicon-based functional nano-enzyme material still has good magnetism and is convenient to recover.
FIG. 5 is a TEM image of the magnetic particle, the magnetic particle-silicon-coated and magnetic silicon-based functional nanoenzyme material of the present embodiment; as can be seen from the graph A, the synthesized magnetic particles have rough surfaces, uniform particle sizes and better dispersibility, which is consistent with the result of FIG. 2; when wrapping SiO2And then, the surface becomes smooth, and after PtAu is further loaded, small particles appear on the smooth particle surface and become rough, so that the successful synthesis of the functional nano-enzyme material is proved.
Example 2
A preparation method of a magnetic silicon-based functional nano enzyme material comprises the following steps:
1. synthesis of gold nanoparticles (AuNPS): taking 1.5mL of 24mM chloroauric acid and 7.3g of trisodium citrate, diluting to 100mL, then placing on a magnetic stirrer, adding 3mLNaBH4(11 mg of the solution was dissolved in 3mL of ice water), and the mixture was stirred at room temperature overnight.
2、Fe3O4Magnetic nanoparticle synthesis: 1.08g of FeCl3And 0.4g of citric acid in 25mL of diethylene glycol, then 2.4g of sodium acetate was added with stirring, stirring was continued for 30min, the mixture was transferred to a reaction vessel, reacted at 180 ℃ for 12h, then cooled to room temperature, and washed 5 times with water and ethanol.
3、Fe3O4Coating silicon dioxide on the surface of the magnetic nano-particles: and (3) ultrasonically dispersing 500 mu L of the synthesized stock solution of the magnetic beads in 5mL of ultrapure water, sucking 27mL of absolute ethyl alcohol by using a liquid-transferring gun, adding the absolute ethyl alcohol into a three-neck flask, after ultrasonic dispersion is carried out for 5min, erecting the three-neck flask and a stirring device at the rotating speed of 300-500 rpm (sufficient stirring and medium speed), adding 125 mu L of concentrated ammonia water (slowly adding), and stirring for 5-10 min. At this time, 60 μ L of TEOS (TEOS: absolute ethanol ═ 1:1) was further added dropwise, and the reaction was stirred at room temperature overnight. After the reaction is finished, the solution is washed with ethanol and ultrapure water for three times, and finally the volume is adjusted to 5mL by water.
4. Amination: taking 500 mu L of stock solution coated with silicon, using ethanol to fix the volume to 5mL, then adding 20 mu L of LAPTES (dissolved in 500 mu L of ethanol), fully mixing, adding 125 mu L of strong ammonia water, stirring at room temperature for reaction for 12h, finishing the reaction, washing with ethanol for 3 times, and fixing the volume to 2 mL.
5. Taking 500 mu L of aminated magnetic nanoparticles, removing ethanol, adding excessive AuNPs, performing ultrasonic treatment for 15min, and performing magnetic separation to remove excessive supernatant. The obtained magnetic particles loaded with nanogold were redispersed in 8mL of ultrapure water, and 200. mu. L H was slowly added thereto with stirring2PtCl6(19.32mM), reacting for 5-20 min, and slowly adding NaBH4(0.01 g of the solution was dissolved in 1mL of ice water), then aged for 12h, and finally subjected to centrifugationCleaning heart, keeping solid, dispersing again and storing.
6. And (3) putting 0.5mL of the prepared functional magnetic nano material into a glass bottle, adding 100 mu L of TMB, standing for 3min, and enabling the mixed solution to show blue, which indicates that the prepared nano enzyme material has the function of catalyzing the TMB to show color and hydrogen peroxide is not added.
Example 3
A preparation method of a magnetic silicon-based functional nano enzyme material comprises the following steps:
1. synthesis of gold nanoparticles (AuNPS). Taking 1.5mL of 24.32mM chloroauric acid and 6.5g of trisodium citrate, diluting to 100mL, then placing on a magnetic stirrer, adding 3mL NaBH4(11 mg of the solution was dissolved in 3mL of ice water), and the mixture was stirred at room temperature overnight.
2、Fe3O4And (3) synthesizing magnetic nanoparticles. 0.6g of FeCl3And 0.3g of trisodium citrate in 20mL of ethylene glycol, then 1.2g of sodium acetate was added under stirring, stirring was continued for 30min, and the mixture was transferred to a reaction vessel, reacted at 180 ℃ for 12h, then cooled to room temperature, and washed 5 times with water and ethanol.
3、Fe3O4The surface of the magnetic nano-particles is coated with silicon dioxide. And (3) ultrasonically dispersing 1mL of the synthesized magnetic bead stock solution in 5mL of ultrapure water, then sucking 25mL of absolute ethyl alcohol by using a liquid-transferring gun, adding the absolute ethyl alcohol into a three-neck flask, after ultrasonically dispersing for 5min, erecting the three-neck flask and a stirring device at the rotating speed of 300-500 rpm (sufficient stirring and medium speed), adding 250 mu L of concentrated ammonia water (slowly adding), and stirring for 5-10 min. At this time, 80 μ L of TEOS (TEOS: absolute ethanol ═ 1:1) was further added dropwise, and the reaction was stirred at room temperature overnight. After the reaction is finished, the solution is washed with ethanol and ultrapure water for three times, and finally the volume is adjusted to 5mL by water.
4. And (4) amination. Taking 500 mu L of stock solution coated with silicon, using ethanol to fix the volume to 5mL, then adding 20 mu L of LAPTES (dissolved in 500 mu L of ethanol), fully mixing, adding 150 mu L of strong ammonia water, stirring at room temperature for reaction for 12h, finishing the reaction, washing with ethanol for 3 times, and fixing the volume to 2 mL.
5. Taking 500 mu L of aminated magnetic nanoparticles, removing ethanol, and adding excessiveAuNPs, ultrasonic for 15min, magnetic separation to remove excess supernatant. The obtained magnetic particles loaded with nanogold were redispersed in 8mL of ultrapure water, and 200. mu. L H was slowly added thereto with stirring2PtCl6(19.32mM), reacting for 5-20 min, and slowly adding NaBH4(0.017 g of the solid is dissolved in 2mL of ice water), then the mixture is aged for 12 hours, and finally the solid is retained after centrifugal cleaning, and the solid is stored after redispersion.
6. And (3) putting 1.5mL of the prepared functional magnetic nano-enzyme material into a glass bottle, adding 150 mu L of TMB, standing for 1min, and enabling the mixed solution to develop blue, which indicates that the prepared nano-enzyme material has the function of catalyzing the TMB to develop color and hydrogen peroxide is not added.
Example 4
The catalytic effect of the magnetic nano enzyme under different pH values is as follows:
the functional magnetic nanoenzyme prepared in example 1 is re-dispersed in 1ml PBS buffer solution with pH values of 3, 4, 5, 6, 7, 8 and 9 from left to right in sequence, and ultrasonically dispersed for 1 min. Then respectively adding 50 mu L of TMB with the same concentration and the same amount, and standing for 1-3 min. The results show that: TMB develops the fastest at pH 3 and pH 4 and eventually turns yellow over time; TMB develops relatively quickly at pH 5 and pH 6; no color development when the pH is 7-9.
Example 5
The magnetic silicon-based functional nano enzyme material has the color development effects on ABTS, OPD and TMB respectively:
dividing 12 PE pipes into 4 groups, respectively adding ABTS, OPD and TMB solution 100uL with the pH value of 4-5 in 4 groups of experiments, and then adding the mixture into a 1 st group: adding H2O2And 200uL of the magnetic silicon-based functional nano enzyme material prepared in the embodiment 1, standing for 1-7 min, and observing the color development effect; group 2: without addition of H2O2However, 200uL of the magnetic silicon-based functional nano enzyme material prepared in the embodiment 1 is added, and the color development effect is observed after standing for 1-7 min; group 3: adding H2O2But the magnetic silicon-based functional nano enzyme material is not added, and the mixture is kept stand for 1-7 min as a reference; group 4: without addition of H2O2And (3) also adding a magnetic silicon-based functional nano enzyme material, and standing for 1-7 min as a control.
As a result: the magnetic silicon-based functional nano enzyme material can develop color for three chromogen reagents. If H is added2O2The color development effect is more obvious, but H is not added2O2Color development is also possible. In addition, the TMB color development effect is most obvious in the three chromogen reagents.
Claims (10)
1. A preparation method of a magnetic silicon-based functional nano enzyme material is characterized by comprising the following preparation steps: synthesis of Nanogold particles, Fe3O4Magnetic nanoparticle synthesis, Fe3O4Coating silicon dioxide on the surface of the magnetic nano-particles, amination and synthesis of a nano-enzyme material;
and (3) synthesis of nano gold particles: taking 0.1-3 mL of 15-30 mM chloroauric acid and 4-10 g of trisodium citrate, diluting to a constant volume of 100mL, then placing on a magnetic stirrer, and adding 0.5-5 mLNaBH4And stirring the solution at normal temperature for 10-12 h.
2. The method for preparing magnetic silicon-based functional nanoenzyme material according to claim 1, wherein the Fe is Fe3O4Magnetic nanoparticle synthesis: 0.5g to 10g of FeCl3And 0.1-2 g of trisodium citrate are dispersed in 20-40 mL of ethylene glycol, then 0.1-3 g of sodium acetate is added under the stirring condition, the mixture is continuously stirred for 20-60 min, the mixture is transferred into a reaction kettle, the reaction is carried out for 8-12 h under the condition of 180-300 ℃, then the reaction kettle is cooled to room temperature, and the mixture is washed with water and ethanol for multiple times.
3. The method for preparing magnetic silicon-based functional nanoenzyme material according to claim 1, wherein the Fe is Fe3O4Coating silicon dioxide on the surface of the magnetic nano-particles: ultrasonically dispersing 100-800 mu L of synthetic magnetic bead stock solution into 5mL of ultrapure water, then sucking 5-35 mL of absolute ethyl alcohol by using a liquid transfer gun, adding into a container, after ultrasonically dispersing for 2-5 min, erecting the container and a stirring device, then adding 100-500 mu L of concentrated ammonia water into the container, and stirring for 5-10 min; at the moment, 10-100 mu L TEOS solution is added dropwise, and the mixture is stirred and reacts for 10-12 h at room temperature; after the reaction is finished, the catalyst is reacted with the catalyst BAnd cleaning the alcohol and the ultrapure water for multiple times, and finally, metering the volume to 1-10 mL by using water.
4. The method for preparing a magnetic silicon-based functional nanoenzyme material according to claim 1, wherein the amination: taking 100-800 mu L of stock solution coated with silicon, using ethanol to fix the volume to 5mL, then adding APTES solution, fully mixing, adding 20-200 mu L of concentrated ammonia water, stirring at room temperature to react for 6-20 h, after the reaction is finished, washing with ethanol for multiple times, and fixing the volume to 2 mL.
5. The method for preparing a magnetic silicon-based functional nanoenzyme material according to claim 1, wherein the synthesis of the nanoenzyme material comprises the following steps: taking 10-800 mu L of aminated magnetic nanoparticles, removing ethanol, adding AuNPs, performing ultrasonic treatment for 15min, performing magnetic separation to obtain a supernatant with transparent color and no Au NPs color, judging that the AuNPs are successfully adsorbed on the surface of the magnetic beads, and repeating the process until the supernatant is wine red in color; re-dispersing the obtained magnetic particles loaded with the nanogold in 2-10 mL of ultrapure water, and slowly adding 50-500 mu L of H with the concentration of 10-25 mM under the stirring condition2PtCl6After reacting for 5-20 min, slowly adding NaBH4And aging the solution for 3-12 h, finally carrying out centrifugal cleaning, retaining the solid, re-dispersing and storing.
6. The method for preparing magnetic silicon-based functional nanoenzyme material according to claim 1, wherein the NaBH is prepared by a method comprising a step of adding NaBH to a solution of the NaBH, and a step of adding NaBH to the solution of the NaBH to obtain the magnetic silicon-based functional nanoenzyme material4The ice cream is prepared by dissolving 5-20 mg of the ice cream in 1-5 mL of ice water.
7. The method for preparing a magnetic silicon-based functional nanoenzyme material according to claim 3, wherein the stirring speed is 300-500 rpm.
8. The method of claim 3, wherein the TEOS solution is prepared by mixing TEOS and absolute ethanol at a ratio of 1: 1.
9. A magnetic silicon-based functional nanoenzyme material prepared by the preparation method according to any one of claims 1 to 8.
10. The magnetic silicon-based functional nano-enzyme material prepared according to any one of claims 1 to 8 is used for color development of 3,3 ', 5, 5' -tetramethylbenzidine, o-phenylenediamine or 2, 2-diaza-bis (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt.
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